1. Field of the Invention
The present invention relates generally to methods for assessment of carcinoma cancer therapy and relapse detection, and more specifically it relates to an efficacy assessment method for lung cancer therapy to rapidly predict the outcome of lung cancer therapy so treatment with higher likelihood of success can be selected to prevent invalid treatment from wrecking patents, and a routine monitoring method for cancer relapse after the treatment.
2. Description of the Related Art
Lung cancer is the leading cause of cancer-related death and non-small cell lung cancer (NSCLC) accounts for ˜80% of the cases. Attempts to use serum protein markers for the early diagnosis of lung cancer have not yielded satisfactory results for routine screening, and newly developed early diagnostic methods using serum DNA as a diagnostic marker await further validation. Current therapeutic measures remain unable to lower the mortality rate of late-stage lung cancer patients. Surgical resection is still the best cure for the early-stage patients. The tumor, node, metastasis (TNM) classification has been used for cancer staging and prognosis for decades. A large portion of early-stage patients, defined by the current staging system and available imaging modalities, still develop distant metastases although they received surgical removal of the tumor mass. The inability to detect disseminated tumor cells with the current imaging techniques is a major obstacle to accurate cancer staging.
NSCLC is heterogeneous with respect to histology and biological characteristics. Individual NSCLC cells within a tumor and in different patients' tumors express different amounts of marker gene transcripts. The heterogeneity of marker gene expression levels in NSCLC cells limits the reliability of an assay method with a single-marker detection scheme. Several literature reports have described PCR methods for the detection of tumor cells dispersed in the circulation. However, not one tumor marker is consistently and specifically expressed in all of the primary tumors of a particular malignancy. Literature reports have also shown that a panel of marker genes provides a more reliable and informative approach than a single-marker assay for the detection of melanoma and breast cancer cells in blood. Such assays for lung cancer have been limited by the availability of molecular markers.
The presence of epithelial cancer cells in the bone marrow and in the peripheral blood of patients with carcinoma has been reported in literature reports and prior arts. In contrast to bone marrow aspirates, peripheral blood samples can be obtained routinely and more readily. Carcinoma accounts for around 85% of human cancers and the carcinoma cells are of epithelial cell lineage. Techniques such as immunocytology and flow cytometry have been employed in prior arts to detect circulating cancer cells in the peripheral blood. However, both techniques are based on extracting or labeling intact carcinoma cells in circulation by antibodies targeting specifically to the epithelial cell surface antigens such as EpCAM and others. Malignant carcinoma cancer cells often are de-differentiated and lose the characteristic epithelial cell surface antigens. In addition, it is known in cancer research field that EpCAM gene expression is often suppressed to facilitate tumor metastasis. Therefore, the antibody based detection methods have been reported to have low positive detection rates or high false negative rates. Polymerase chain reaction (PCR) has been employed to detect disseminated tumor cells in peripheral blood. Several literature reports have described the use of PCR for detecting circulating cancer cells in the peripheral blood of patients of various cancers. For instance, Peck et al., reported the use of cytokeratin 19 as the maker gene for detecting circulating cancer cells in NSCLC patients with an overall positive detection rate around 40%.
Compared with immunocytology and flow cytometry, PCR has the advantages that it is more readily available, less involved in the operating procedures, less instrument cost, and others. On the other hand, PCR is not able to yield the number of counts of circulating cancer cell in a sample like the other two techniques.
To overcome the current technology difficulties in achieving high positive detection rate and rapid assessment of lung cancer therapy efficacy and relapse detection, a panel of marker genes for achieving high positive detection rate by qPCR and a quantitative analysis method for predicting lung cancer treatment outcome and for prognosis are needed.